货号: | 0870112DK |
数量: | 100 |
保存条件: | 2-8 |
规格: | 1000ml/瓶 |
Intended Use For human ex-vivo tissue and cell culture processing applications. CAUTION: When used as a medical device, Federal Law restricts this device to sale by or on the order of a physician. Safety Information For every chemical, read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Human origin materials are non-reactive (donor level) for anti-HIV 1 & 2, anti-HCV and HBsAg. Handle in accordance with established bio-safety practices. Use AIM V® Medium CTS™ Serum Free Medium comes supplemented with L-glutamine, streptomycin sulfate, and gentamicin sulfate. Additional supplementation with cytokines or growth factors may be required per specific investigator’s procedures and should be aseptically added immediately prior to use.
Cell Culture The following protocol serves as a general guideline for static T cell and dendritic cell culture, regardless of vessel. Feed and maintain cells at desired concentrations while cells are in log phase growth. Dilute cells to a viable cell density of 5 × 105 cells/mL whenever the viable cell density reaches ≥1 × 106 cells/mL. For optimal gas exchange in static plate cultures it is recommended that medium depth not exceed 1–1.2 cm. For highdensity culture in bioreactors, optimal procedures should be determined empirically by the investigator. T Cell Culture 1. Prepare fresh peripheral blood mononuclear cells (PBMCs) or rapidly thaw (<1 minute) a vial of frozen PBMCs in a 37°C water bath according to standard PBMC thawing protocols. 2. Wash cells with Dulbecco's Phosphate Buffered Saline (DPBS) CTS™ without calcium and magnesium, supplemented with 2–5% heat-inactivated human pooled Type AB serum according to the application, if desired or required. 3. Determine viable cell density using a Countess® Automated Cell Counter. 4. Centrifuge cells at 200 × g for 5–10 minutes and aspirate wash buffer supernatant. 5. Resuspend PBMC pellet at approximately 0.5–1 × 106 cells/mL in medium supplemented with cytokines (e.g., IL-2), if used at culture initiation. Transfer the desired number of cells to the desired tissue culture vessel. Note: A variety of protocols may be used for activating T cells for subsequent expansion, including adding stimulatory antibodies or antigen presenting cells. Similarly, for either small or large scale T cell expansion, cells can be isolated, activated and expanded using Dynabeads® CD3/CD28 CTS™ according to instructions in the product insert.